Carbonate clumped isotopes offer a potentially transformational tool to interpret Earth's history, but the proxy is still limited by poor interlaboratory reproducibility. Here, we focus on the ...uncertainties that result from the analysis of only a few replicate measurements to understand the extent to which unconstrained errors affect calibration relationships and paleoclimate reconstructions. We find that highly precise data can be routinely obtained with multiple replicate analyses, but this is not always done in many laboratories. For instance, using published estimates of external reproducibilities we find that typical clumped isotope measurements (three replicate analyses) have margins of error at the 95% confidence level (CL) that are too large for many applications. These errors, however, can be systematically reduced with more replicate measurements. Second, using a Monte Carlo‐type simulation we demonstrate that the degree of disagreement on published calibration slopes is about what we should expect considering the precision of Δ47 data, the number of samples and replicate analyses, and the temperature range covered in published calibrations. Finally, we show that the way errors are typically reported in clumped isotope data can be problematic and lead to the impression that data are more precise than warranted. We recommend that uncertainties in Δ47 data should no longer be reported as the standard error of a few replicate measurements. Instead, uncertainties should be reported as margins of error at a specified confidence level (e.g., 68% or 95% CL). These error bars are a more realistic indication of the reliability of a measurement.
Key Points
Highly precise Δ47 data can be obtained with multiple replicate analyses
Disagreements in calibration slopes can be largely explained by poor sample replication, small sample sizes, and narrow temperature ranges
Uncertainties on Δ47 data should be reported as margins of error at a specified confidence level (e.g., 68% or 95% CL)
Chemotherapy is still the cornerstone for malaria control. Developing drugs against Plasmodium parasites and monitoring their efficacy requires methods to accurately determine the parasite killing ...rate in response to treatment. Commonly used techniques essentially measure metabolic activity as a proxy for parasite viability. However, these approaches are susceptible to artefacts, as viability and metabolism are two parameters that are coupled during the parasite life cycle but can be differentially affected in response to drug actions. Moreover, traditional techniques do not allow to measure the speed-of-action of compounds on parasite viability, which is an essential efficacy determinant. We present here a comprehensive methodology to measure in vitro the direct effect of antimalarial compounds over the parasite viability, which is based on limiting serial dilution of treated parasites and re-growth monitoring. This methodology allows to precisely determine the killing rate of antimalarial compounds, which can be quantified by the parasite reduction ratio and parasite clearance time, which are key mode-of-action parameters. Importantly, we demonstrate that this technique readily permits to determine compound killing activities that might be otherwise missed by traditional, metabolism-based techniques. The analysis of a large set of antimalarial drugs reveals that this viability-based assay allows to discriminate compounds based on their antimalarial mode-of-action. This approach has been adapted to perform medium throughput screening, facilitating the identification of fast-acting antimalarial compounds, which are crucially needed for the control and possibly the eradication of malaria.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Aims
The PREVIEW lifestyle intervention study (ClinicalTrials.gov Identifier: NCT01777893) is, to date, the largest, multinational study concerning prevention of type‐2 diabetes. We hypothesized that ...the initial, fixed low‐energy diet (LED) would induce different metabolic outcomes in men vs women.
Materials and methods
All participants followed a LED (3.4 MJ/810 kcal/daily) for 8 weeks (Cambridge Weight Plan). Participants were recruited from 8 sites in Europe, Australia and New Zealand. Those eligible for inclusion were overweight (BMI ≥ 25 kg/m2) individuals with pre‐diabetes according to ADA‐criteria. Outcomes of interest included changes in insulin resistance, fat mass (FM), fat‐free mass (FFM) and metabolic syndrome Z‐score.
Results
In total, 2224 individuals (1504 women, 720 men) attended the baseline visit and 2020 (90.8%) completed the follow‐up visit. Following the LED, weight loss was 16% greater in men than in women (11.8% vs 10.3%, respectively) but improvements in insulin resistance were similar. HOMA‐IR decreased by 1.50 ± 0.15 in men and by 1.35 ± 0.15 in women (ns). After adjusting for differences in weight loss, men had larger reductions in metabolic syndrome Z‐score, C‐peptide, FM and heart rate, while women had larger reductions in HDL cholesterol, FFM, hip circumference and pulse pressure. Following the LED, 35% of participants of both genders had reverted to normo‐glycaemia.
Conclusions
An 8‐week LED induced different effects in women than in men. These findings are clinically important and suggest gender‐specific changes after weight loss. It is important to investigate whether the greater decreases in FFM, hip circumference and HDL cholesterol in women after rapid weight loss compromise weight loss maintenance and future cardiovascular health.
Malaria is a devastating infection caused by protozoa of the genus Plasmodium. Drug resistance is widespread, no new chemical class of antimalarials has been introduced into clinical practice since ...1996 and there is a recent rise of parasite strains with reduced sensitivity to the newest drugs. We screened nearly 2 million compounds in GlaxoSmithKline's chemical library for inhibitors of P. falciparum, of which 13,533 were confirmed to inhibit parasite growth by at least 80% at 2 microM concentration. More than 8,000 also showed potent activity against the multidrug resistant strain Dd2. Most (82%) compounds originate from internal company projects and are new to the malaria community. Analyses using historic assay data suggest several novel mechanisms of antimalarial action, such as inhibition of protein kinases and host-pathogen interaction related targets. Chemical structures and associated data are hereby made public to encourage additional drug lead identification efforts and further research into this disease.
Celotno besedilo
Dostopno za:
DOBA, IJS, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Benzoxaboroles are effective against bacterial, fungal and protozoan pathogens. We report potent activity of the benzoxaborole AN3661 against Plasmodium falciparum laboratory-adapted strains (mean IC
...32 nM), Ugandan field isolates (mean ex vivo IC
64 nM), and murine P. berghei and P. falciparum infections (day 4 ED
0.34 and 0.57 mg kg
, respectively). Multiple P. falciparum lines selected in vitro for resistance to AN3661 harboured point mutations in pfcpsf3, which encodes a homologue of mammalian cleavage and polyadenylation specificity factor subunit 3 (CPSF-73 or CPSF3). CRISPR-Cas9-mediated introduction of pfcpsf3 mutations into parental lines recapitulated AN3661 resistance. PfCPSF3 homology models placed these mutations in the active site, where AN3661 is predicted to bind. Transcripts for three trophozoite-expressed genes were lost in AN3661-treated trophozoites, which was not observed in parasites selected or engineered for AN3661 resistance. Our results identify the pre-mRNA processing factor PfCPSF3 as a promising antimalarial drug target.
Pericytes (PC) are mural cells that surround endothelial cells in small blood vessels. PC have traditionally been credited with structural functions, being essential for vessel maturation and ...stabilization. However, an accumulating body of evidence suggests that PC also display immune properties. They can respond to a series of pro-inflammatory stimuli and are able to sense different types of danger due to their expression of functional pattern-recognition receptors, contributing to the onset of innate immune responses. In this context, PC not only secrete a variety of chemokines but also overexpress adhesion molecules such as ICAM-1 and VCAM-1 involved in the control of immune cell trafficking across vessel walls. In addition to their role in innate immunity, PC are involved in adaptive immunity. It has been reported that interaction with PC anergizes T cells, which is attributed, at least in part, to the expression of PD-L1. As components of the tumor microenvironment, PC can also modulate the antitumor immune response. However, their role is complex, and further studies will be required to better understand the crosstalk of PC with immune cells in order to consider them as potential therapeutic targets. In any case, PC will be looked at with new eyes by immunologists from now on.
Malaria is one of the most significant causes of childhood mortality, but disease control efforts are threatened by resistance of the Plasmodium parasite to current therapies. Continued progress in ...combating malaria requires development of new, easy to administer drug combinations with broad-ranging activity against all manifestations of the disease. DSM265, a triazolopyrimidine-based inhibitor of the pyrimidine biosynthetic enzyme dihydroorotate dehydrogenase (DHODH), is the first DHODH inhibitor to reach clinical development for treatment of malaria. We describe studies profiling the biological activity, pharmacological and pharmacokinetic properties, and safety of DSM265, which supported its advancement to human trials. DSM265 is highly selective toward DHODH of the malaria parasite Plasmodium, efficacious against both blood and liver stages of P. falciparum, and active against drug-resistant parasite isolates. Favorable pharmacokinetic properties of DSM265 are predicted to provide therapeutic concentrations for more than 8 days after a single oral dose in the range of 200 to 400 mg. DSM265 was well tolerated in repeat-dose and cardiovascular safety studies in mice and dogs, was not mutagenic, and was inactive against panels of human enzymes/receptors. The excellent safety profile, blood- and liver-stage activity, and predicted long half-life in humans position DSM265 as a new potential drug combination partner for either single-dose treatment or once-weekly chemoprevention. DSM265 has advantages over current treatment options that are dosed daily or are inactive against the parasite liver stage.
The eastern Mediterranean Sea lies under the influence of high- and low-latitude climatic systems. The northern part of the basin is affected by Atlantic depressions and continental and polar air ...masses that promote intermediate and deep-water formation. The southern part is influenced by subtropical conditions and monsoon activity. Monsoon intensification results in enhanced freshwater discharge from the Nile River and other (now dry) systems along the North African margin. This freshwater influx into the Mediterranean Sea reduces surface water buoyancy loss. Disentangling the influences of these diverse climatic forcings is hindered by inherent proxy data limitations and by interactions between the climatic forcings. Here we use a wealth of published and new paleoclimate records across Termination II to understand the impacts of the higher latitude and subtropical/monsoon climate influences on coccolithophore ecology and holococcolith preservation in Aegean Sea sediment core LC21. We then use these findings to interpret coccolith assemblage variations at Ocean Drilling Program Site 967 (located nearby LC21, at the Eratosthenes Seamount) during multiple glacial-interglacial cycles across the Middle Pleistocene (marine isotopic stages 14–9). The LC21 analysis suggests that holococcolith preservation was enhanced during Heinrich Stadial 11 (∼133 ka) and cold spell C26 (∼119 ka). These two events have been previously linked to cold conditions in the North Atlantic and Atlantic Meridional Overturning Circulation weakening. We propose that associated atmospheric perturbations over the Mediterranean Sea promoted deep-water formation, and thus holococcolith preservation. Similarly, in the Middle Pleistocene (MIS 14-9) of Site 967, we observe temporal coincidence between ten episodes of enhanced holococcolith preservation and episodes of Atlantic Meridional Overturning Circulation slowdown. In Site 967, we also identified repeated fluctuations in placoliths and in Florisphaera profunda, which indicate nutricline depth variations. The development of a deep chlorophyll maximum is associated with the North Africa and wet phases, as recently observed using elemental proxy records at Site 967, during the deposition of sapropel layers. A further deep chlorophyll maximum development is identified during MISs 12 and 10, as a result of pycnocline and nutricline shoaling within the lower part of the photic zone due to glacial sea-level lowering and water mass transport reduction at both the Gibraltar and Sicily Straits. Finally, enhanced holococcolith preservation during cold/dry events is clearly correlated to weakened monsoon activity in both Africa and Asia.
•F. profunda in sapropel S5 is in line with maximum rates of δ18O Asian speleothems and with the CH4 overshoot in EDC;•F. profunda DCM development in glacials, due to sea-level lowering that reduced water-mass transport through straits;•Holococcolith enhanced preservation during cold spells, in response to AMOC slowdown and weakened monsoon activity;
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